猪源SIRT5促进O型口蹄疫病毒在PK-15细胞复制  

Porcine SIRT5 promotes replication of foot and mouth disease virus type O in PK-15 cells

作　　者：陈国辉 史喜绢 别鑫恬 杨行 赵思越 张大俊 赵登率 闫文倩 陈玲玲 赵美玉 何路 郑海学[2] 刘霞[1] 张克山[2] CHEN Guo-hui;SHI Xi-juan;BIE Xin-tian;YANG Xing;ZHAO Si-yue;ZHANG Da-jun;ZHAO Deng-shuai;YAN Wen-qian;CHEN Ling-ling;ZHAO Mei-yu;HE Lu;ZHENG Hai-xue;LIU Xia;ZHANG Ke-shan(College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences/College of Veterinary Medicine of Lanzhou University/State Key Laboratory for the Prevention and Control of Animal Diseases,Lanzhou 730046,China)

机构地区：[1]甘肃农业大学生命科学技术学院,兰州730070 [2]中国农业科学院兰州兽医研究所,兰州大学动物医学与生物安全学院动物疫病防控国家重点实验室,兰州730046

出　　处：《中国人兽共患病学报》2024年第5期421-429,共9页Chinese Journal of Zoonoses

基　　金：国家重点研发计划项目(No.2021YFD1800300);甘肃省科技重大专项(No.21ZD3NA001-5);中国农业科学院重大任务(No.CAASZDRW202006-03)联合资助。

摘　　要：目的探究猪源SIRT5对O型口蹄疫病毒(Foot and mouth disease virus serotype O,FMDV-O)复制的影响及其调控FMDV-O复制的初步机制。方法利用Western Blotting和RT-qPCR检测FMDV-O感染PK-15细胞后内源性SIRT5表达情况;设计合成3对SIRT5特异性siRNA,通过Western Blotting和RT-qPCR检测SIRT5、FMDV-O蛋白水平、转录水平及病毒拷贝数的变化情况;构建SIRT5真核表达质粒转染至PK-15细胞,运用Western Blotting、RT-qPCR实验方法探究过表达SIRT5对FMDV-O复制的影响,同时利用RT-qPCR检测过表达SIRT5对SeV、FMDV-O诱导的I型干扰素刺激基因mRNA表达水平的影响。结果FMDV-O感染PK-15细胞后内源性SIRT5的表达上调;siRNA干扰SIRT5抑制FMDV-O的复制;过表达SIRT5促进FMDV-O复制;过表达SIRT5降低了SeV、FMDV-O诱导的干扰素刺激基因mRNA的表达水平。结论FMDV-O感染刺激宿主SIRT5表达,而猪源SIRT5通过抑制I型干扰素刺激基因的产生进而促进FMDV-O复制。本研究为进一步探究猪源SIRT5促进FMDV-O复制的机制提供参考依据。The effect of porcine SIRT5 on replication of foot and mouth disease virus type O(FMDV-O) and the underlying regulatory mechanism were investigated.Western blot and RT-qPCR analyses were employed to monitor expression of endogenous SIRT5 in PK-15 cells infected with FMDV-O.Three pairs of SIRT5-specific siRNAs were synthesized.Changes to SIRT5 and FMDV-O protein and transcript levels,in addition to virus copy numbers,were measured by western blot and RT-qPCR analyses.PK-15 cells were transfected with a eukaryotic SIRT5 expression plasmid.Western blot and RT-qPCR analyses were used to explore the impact of SIRT5 overexpression on FMDV-O replication.Meanwhile,RT-qPCR analysis was used to detect the effect of SIRT5 overexpression on the mRNA expression levels of type I interferon-stimulated genes induced by SeV and FMDV-O.The results showed that expression of SIRT5 was up-regulated in PK-15 cells infected with FMDV-O and siRNA interfered with SIRT5 to inhibit FMDV-O replication.SIRT5 overexpression promoted FMDV-O replication.SIRT5 overexpression decreased mRNA expression levels of interferon-stimulated genes induced by SeV and FMDV-O.These results suggest that FMDV-O infection stimulated expression of SIRT5 in PK-15 cells,while SIRT5 promoted FMDV-O replication by inhibiting production of type I interferon-stimulated genes.These findings provide a reference to further explore the mechanism underlying the ability of porcine SIRT5 to promote FMDV-O replication.

关 键 词：猪源SIRT5 FMDV-O 干扰素刺激基因 PK-15细胞 

分 类 号：R383.3[医药卫生—医学寄生虫学]